LOCUS       V041309                 7161 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041309
VERSION     V041309
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1841
                     /label="Ciart(NM_001134593)"
                     /note="Ciart(NM_001134593)"
                     /gene="Ciart"
     CDS             1848..1913
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1937..2525
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2591..2639
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2841..3269
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3283..3612
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3679..4473
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4649..4770
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4819..4835)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4843..4859
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(4867..4897)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4912..4933
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5221..5809)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5980..6840)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(6841..6945)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga ttctccatct agcgtttctt
      781 cttattcctc ctcctctctc tccccgtcat tttccacgtc ccctgtgaac agtgacttta
      841 gcttcccatc tgataatgag agggagggca tgcgcacccg tgagctcagg ccagacactg
      901 tggggcagag aggaggttcc aggcccagtc caggccccat ccgctgccga caccgaccca
      961 gggtttctag taaccaacat acagcatctc acctggaaca gcaaggatct gaagtcaaaa
     1021 gatcaagaga cggtgaattg gagacaagtc taaacaccca gggatgtact acagagggag
     1081 acctgctctt tgctcaaaag tgtaaagaac tccaagggtt catacggccc ctcacagacc
     1141 tactgaacgg attaaaaatg ggtcgctttg acagaggatt aagtagcttc caacagagcg
     1201 tggccatgga ccggatccag cgtatagtgg gtgttttaca gaagcctcat atgggagagc
     1261 gttatctagg aaccctgctg caggtggaag gcatgctaaa gacttggttt cctcatatag
     1321 ctgcccagaa atcatcctta ggaggaagca ggcatcagat aagcaagcac ttccctagcc
     1381 accacagtga ttcaggtgct tcttctcctg cgcctctcat agggaagatg ggccagacac
     1441 agctgggaca tttagttttg aaaccaaagc agccttggca tctcacagga tggccggcta
     1501 tgaacctcac ctggatccat gacactccaa tttgtaaccc tcccctcggt tcccaaggct
     1561 ccacctctgg ccacagcccc aacggcactg gagccagcat tggcgtcatc cttgtcctcc
     1621 agaaaggagg acagcccttc acccattctg ccccgggcac tccaatccca ccaactacac
     1681 tgtctcctgt cctccctggt gatctgagga aactgtcggg agaggagcct cgctgccaca
     1741 gtttgccagt aacgctgcca tcagactgga ggtgcatcct ttgtccccct gtcctaccta
     1801 ccgcagatcg agaggtgact acaggacacc agagccacag aggatccgac tacaaagacc
     1861 atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac aagtagtgag
     1921 ggcccgatat ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1981 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     2041 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     2101 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     2161 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     2221 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     2281 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     2341 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     2401 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     2461 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     2521 cccgcctgga aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     2581 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     2641 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     2701 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     2761 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg
     2821 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2881 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2941 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat
     3001 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3061 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3181 ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga
     3241 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3361 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3421 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3481 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3541 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3601 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     3661 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3721 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3781 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3841 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     3901 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3961 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4021 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4081 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4141 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4201 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4261 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4321 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4381 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4441 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac
     4501 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4561 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4621 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4681 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4741 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     4801 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     4861 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     4921 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5401 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5641 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5821 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6961 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt
     7021 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7081 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7141 tgcatgaaga atctgcttag g
//
